Abstract Epigenetic variation in natural populations with contrasting habitats might be an important element, in addition to the genetic variation, in plant adaptation to environmental stress. Here, we assessed genetic, epigenetic, and cytogenetic structure of the three Lilium bosniacum populations growing on distinct habitats. One population was growing under habitual ecological conditions for this species and the other two were growing under stress associated with high altitude and serpentine soil. Amplified fragment length polymorphism and methylation-sensitive amplification polymorphism analyses revealed that the three populations did not differentiate genetically, but were clearly separated in three distinct clusters according to DNA methylation profiles. Principal coordinate analysis showed that overall epigenetic variation was closely related to habitat conditions. A new methylation-sensitive amplification polymorphism scoring approach allowed identification of mainly unmethylated (φST = 0.190) and fully CpG methylated (φST = 0.118) subepiloci playing a role in overall population differentiation, in comparison with hemimethylated sites (φST = 0.073). In addition, unusual rDNA repatterning and the presence of B chromosomes bearing 5S rDNA loci were recorded in the population growing on serpentine soil, suggesting dynamic chromosome rearrangements probably linked to global genome demethylation, which might have reactivated some mobile elements. We discuss our results considering our earlier data on morphology and leaf anatomy of several L. bosniacum populations, and suggest a possible role of epigenetics as a key element in population differentiation associated with environmental stress in these particular lily populations.

The historical changes in European Black Pine population size across the whole natural distribution in Europe and Asia Minor were analyzed facing the Plio-Pleistocene climatic fluctuations. Thirteen chloroplast SSRs and SNPs markers have been studied under the assumptions of “neutral evolution.” Populations and meta-populations had different histories of migration routes, and they were strongly affected by complex patterns of isolation, fragmentation, speciation, expansion (1.88–4.28 Ma), purification selection (2.09–21.41 Ma) and bottleneck (1.85–21.76 Ma). A significant number of populations (min. 29–41%) were in equilibrium for very long periods. Generally, the bottleneck revealed by chloroplast DNA is weaker than the bottleneck revealed by nuclear DNA. The Ne immediately after the bottleneck reaches between 1820 and 3640 individuals. Generally, the historical effective population sizes shrink significantly for the Tertiary period from 10–15 up to 2.5 Ma in Western Europe (by 82%), followed by Asia Minor (69%) and the Balkan Peninsula (28%), likely resulting from important climatic changes. The rates and frequencies of stepwise westwards migration waves have been not sufficient to prevent isolation between the meta-populations and to suppress “sympatric speciation.” The migration was weak for the Pliocene, but was maximal for the Pleistocene, and finally silent for the present interglacial period, namely the Holocene.

The historical changes in European Black Pine population size across the whole natural distribution in Europe and Asia Minor were analyzed facing the Plio-Pleistocene climatic fluctuations. Thirteen chloroplast SSRs and SNPs markers have been studied under the assumptions of “neutral evolution.” Populations and meta-populations had different histories of migration routes, and they were strongly affected by complex patterns of isolation, fragmentation, speciation, expansion (1.88–4.28 Ma), purification selection (2.09–21.41 Ma) and bottleneck (1.85–21.76 Ma). A significant number of populations (min. 29–41%) were in equilibrium for very long periods. Generally, the bottleneck revealed by chloroplast DNA is weaker than the bottleneck revealed by nuclear DNA. The Ne immediately after the bottleneck reaches between 1820 and 3640 individuals. Generally, the historical effective population sizes shrink significantly for the Tertiary period from 10–15 up to 2.5 Ma in Western Europe (by 82%), followed by Asia Minor (69%) and the Balkan Peninsula (28%), likely resulting from important climatic changes. The rates and frequencies of stepwise westwards migration waves have been not sufficient to prevent isolation between the meta-populations and to suppress “sympatric speciation.” The migration was weak for the Pliocene, but was maximal for the Pleistocene, and finally silent for the present interglacial period, namely the Holocene.

Abstract Types and distribution of trichomes, chemical composition of the essential oil, as well as contents of phenolic compounds were studied in four Balcanic Clinopodium L. species (family Lamiaceae), namely C. dalmaticum (Benth.) Brauchler et Heubl, C . pulegium (Rochel) Brauchler, C . serpyllifolium (M. Bieb.) Kuntze, and C . thymifolium (Scop.) Kuntze. Non-glandular trichomes, peltate trichomes, and three types of capitate trichomes were observed on the leaves, calyces, and stems. In general, 62 different essential oil compounds were identified: 21 from C . dalmaticum , 31 from C. thymifolium , 36 from C. serpyllifolium , and 37 from C. pulegium . The essential oil of all species was characterized by the high content of oxygenated monoterpenes (66.2–79.6%), with dominant menthane type compounds, such as menthone, pulegone, piperitenone, and piperitenone oxide. Total polyphenols (TP), tannins (T), total flavonoids (TF), and total phenolic acids (TPA) were determined spectrophotometrically. The lowest levels of these compounds were found in C. serpyllifolium (3.83%, 0.88%, 0.39%, and 2.91%, respectivelly), and the highest in C. thymifolium (TP, 7.40%) and C. pulegium (T, 1.78%; TF, 0.57%; TPA, 6.59%).

Micromeria longipedunculata Bräuchler (syn. M. parviflora (Vis.) Rchb.) is an endemic Illyric-Balkan plant species from Bosnia and Herzegovina, Montenegro, and Albania. We investigated types and distribution of trichomes, pollen morphology, and chemical composition of essential oil (analysed by GC and GC-MS) in M. longipedunculata. Non-glandular trichomes, peltate trichomes, and two types of capitate trichomes (type 1 composed of one basal epidermal cell, and one head cell with subcuticular space; type 2 composed of one basal epidermal cell, two or three stalk cells, and one head cell with subcuticular space) were observed on leaves, bracteoles, the calyx, corolla, and the stem. The pollen grains had six apertures which were set in the equatorial pollen belt and showed medium reticulate ornamentation. A phytochemical analysis of essential oils from four different localities is characterized by similar chemical composition with spathulenol (23.7–39.5%), piperitone oxide (7.7–12.1%) and piperitone (7.3–8.9%) as the major compounds.